Carbonic Anhydrase IX/CA9 Antibody (66.4.C2 (PN-15)) - Azide and BSA Free

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-33152

Novus Biologicals, part of Bio-Techne

Key Product Details

Species Reactivity

Human, Equine

Applications

Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry-Paraffin, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2b Kappa Clone # 66.4.C2 (PN-15)

Format

Azide and BSA Free

Concentration

1.0 mg/ml

View all Carbonic Anhydrase IX/CA9 Antibodies »

Product Specifications

Immunogen

Microsomal fraction of human renal cortical tissue homogenate

Localization

Cell surface and cytoplasmic

Specificity

Recognizes a glycoprotein of ~200kDa, identified as carbonic anhydrase IX (CAIX/gp200). Its epitope resides in the carbohydrate domain of gp200. It shows no significant cross-reactivity with other carbohydrate determinants, such as the Lewis blood group antigens, epithelial membrane antigen, HMFG, and AB blood group antigens. In normal kidney, gp200 is localized along the brush border of the pars convoluta and pars recta segments of the proximal tubule, as well as focally along the luminal surface of Bowmans capsule adjoining the outgoing proximal tubule. Reportedly, gp200 is expressed by 93% of primary and 84% of metastatic renal cell carcinomas. This monoclonal antibody may be useful in the investigations of carcinomas of proximal nephrogenic differentiation especially those showing tubular differentiation.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2b Kappa

Description

1.0 mg/ml of antibody purified from Bioreactor Concentrate by Protein A/G. Prepared in 10mM PBS WITHOUT BSA & azide. Also available at 200 ug/ml WITH BSA & azide (NBP2-15199).

Antibody with azide - store at 2 to 8C. Antibody without azide - store at -20 to -80C.

Scientific Data Images

Immunohistochemistry-Paraffin: Carbonic Anhydrase IX/CA9 Antibody (66.4.C2 (PN-15)) - Azide and BSA Free [NBP2-33152]

Immunohistochemistry-Paraffin: Carbonic Anhydrase IX/CA9 Antibody (66.4.C2 (PN-15)) - Azide and BSA Free [NBP2-33152]

Immunohistochemistry-Paraffin: Carbonic Anhydrase IX/CA9 Antibody (66.4.C2 (PN-15)) - Azide and BSA Free [NBP2-33152] - FFPE human renal cell carcinoma stained with RCC Ab (66.4.C2). Note cytoplasmic & cell surface staining of tumor cells.

Applications

Application
Recommended Usage

Flow Cytometry

Optimal dilutions of this antibody should be experimentally determined.

Immunocytochemistry/ Immunofluorescence

Optimal dilutions of this antibody should be experimentally determined.

Immunohistochemistry-Paraffin

Optimal dilutions of this antibody should be experimentally determined.

Western Blot

Optimal dilutions of this antibody should be experimentally determined.
Application Notes
Immunohistochemistry (Formalin-fixed): 1-2ug/ml for 30 minutes at RT. Staining of formalin-fixed tissues requires heating tissue sections in 10mM Tris with 1mM EDTA, pH 9.0, for 45 min at 95C followed by cooling at RT for 20 minutes.
Optimal dilution for a specific application should be determined.
Trypsin (10 min at 37C) is recommend for antigen retrieval (Bayder & Aydin, 2008).

Formulation, Preparation, and Storage

Purification

Protein A or G purified

Formulation

10 mM PBS

Format

Azide and BSA Free

Preservative

No Preservative

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20 to -80C. Avoid freeze-thaw cycles.

Background: Carbonic Anhydrase IX/CA9

Carbonic anhydrase alpha, isozyme IX, belongs to a family of zinc-containing metalloproteins which hydrate carbon dioxide to generate bicarbonate ions and protons (1). This main catalytic function allows carbonic anhydrase IX to participate in cellular pH regulation. The large family of carbonic anhydrase metalloproteins includes three major classes which have been identified based on sequence and structure analysis. The alpha class is a monomer found in mammals. The beta class may occur as a dimer, tetramer, hexamer or octamer and is found in plants, algae, and bacteria. Lastly, the gamma class is a trimer found in bacteria and represents the most ancient carbonic anhydrase. These three classes of carbonic anhydrase enzymes lack sequence or structural similarities, but all share a conserved active site zinc atom (1).

Carbonic anhydrase IX (theoretical molecular weight 50kDa) belongs to the monomeric alpha class and is a single pass-transmembrane protein with two extracellular domains which serve catalytic and cell adhesion functions (2, 3). By cooperating with sodium bicarbonate cotransporters (NBC), lactate and proton exporting monocarboxylic acid transporters (MCT), and a sodium/hydrogen exchanger (NHE), carbonic anhydrase IX is involved in pH regulation across the cell membrane. This functional property protects cancer cells from intracellular acidification and partly explains the role of carbonic anhydrase IX in cancer cell survival and proliferation. In contrast, the pH regulating activity of carbonic anhydrase IX induces extracellular acidification, which has been implicated in epithelial to mesenchymal transition (EMT) and promoting cancer invasion. Carbonic anhydrase IX is frequently overexpressed in cancer cells (e.g., colorectal-, breast-, lung-carcinoma and brain tumors), an effect promoted by hypoxia within the tumor microenvironment (4). An exception are tumors carrying pVHL inactivating mutations, such as clear cell renal cell carcinoma (ccRCC), where HIF-alpha is stabilized due to dysfunctional proteasomal targeting and can induce HRE (Hypoxia Response Element) containing genes even under physiological normoxia (5). Carbonic anhydrase IX may be detected by immunostaining in tumors, which is found in association with necrotic tissue and metastatic cells. Because the expression of carbonic anhydrase IX correlates with both tumor grade and stage, analysis of its expression in tumors serves as a prognostic factor (4, 6).

References

1. Tripp, B. C., Smith, K., & Ferry, J. G. (2001). Carbonic Anhydrase: New Insights for an Ancient Enzyme. Journal of Biological Chemistry. https://doi.org/10.1074/jbc.R100045200

2. Nishimori, I., & Onishi, S. (2001). Carbonic anhydrase isozymes in the human pancreas. Digestive and Liver Disease. https://doi.org/10.1016/s1590-8658(01)80138-9

3. Zavadova, Z., & Zavada, J. (2005). Carbonic anhydrase IX (CA IX) mediates tumor cell interactions with microenvironment. Oncology Reports. https://doi.org/10.3892/or.13.5.977

4. Pastorekova, S., & Gillies, R. J. (2019). The role of carbonic anhydrase IX in cancer development: links to hypoxia, acidosis, and beyond. Cancer and Metastasis Reviews. https://doi.org/10.1007/s10555-019-09799-0

5. Haase, V. (2009). The VHL Tumor Suppressor: Master Regulator of HIF. Current Pharmaceutical Design. https://doi.org/10.2174/138161209789649394

6. Young, J. R., Coy, H., Kim, H. J., Douek, M., Sisk, A., Pantuck, A. J., & Raman, S. S. (2018). Association of the gross appearance of intratumoral vascularity at MDCT with the carbonic anhydrase IX score in clear cell renal cell carcinoma. American Journal of Roentgenology. https://doi.org/10.2214/AJR.18.19725

Alternate Names

CA9, G250, MN, RCC

Gene Symbol

CA9

Additional Carbonic Anhydrase IX/CA9 Products

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Certificate of Analysis

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Product Specific Notices

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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